Revista Brasileira de Ciência Avícola ISSN: 1516-635X [email protected] Fundação APINCO de Ciência e Tecnologia Avícolas Brasil

Boakye, OD; Emikpe, BO; Folitse, RD; Bonnah, SG; Adusei, K; Owusu, M; Oyebanji, VO Serological Detection of Newcastle Disease Virus Antibodies in Local Chickens and Guinea Fowls in the Area of , Revista Brasileira de Ciência Avícola, vol. 18, núm. 1, enero-marzo, 2016, pp. 87-91 Fundação APINCO de Ciência e Tecnologia Avícolas Campinas, Brasil

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How to cite Complete issue Scientific Information System More information about this article Network of Scientific Journals from Latin America, the Caribbean, Spain and Portugal Journal's homepage in redalyc.org Non-profit academic project, developed under the open access initiative Brazilian Journal of Poultry Science Revista Brasileira de Ciência Avícola Serological Detection of Newcastle Disease Virus ISSN 1516-635X Jan - Mar 2016 / v.18 / n.1 / 087-092 Antibodies in Local Chickens and Guinea Fowls in the http://dx.doi.org/10.1590/18069061-2015-0132 Area of Kumasi, Ghana

Author(s) ABSTRACT

Boakye ODI Newcastle Disease (ND) has been identified as a major constraint to Emikpe BOIV local poultry production with its impact being felt more in rural poultry Folitse RDI Bonnah SGII production which forms about 80% of Ghana poultry population. Adusei KIII However documented evidence on ND virus activity in rural poultry in Owusu MI Oyebanji VOIV Ghana is still lacking. Hence, this study was conducted to evaluate the level of circulating antibodies against ND using the Haemagglutination Inhibition (HI) technique. Sera collected from unvaccinated 292 chickens I School of Veterinary Medicine, Kwame Nkrumah University of Science and Techno- and 153 guinea fowls randomly selected from households and a live bird logy market in Kumasi and its environs were evaluated for Newcastle disease II Ministry of Food and Agriculture, Veterinary Service Department virus antibodies. Results showed 81.8 % (239/292) of local chickens III Veterinary Investigations Laboratory, Kuma- and 24.2 % (37/153) of guinea fowls tested positive for ND antibodies. si. Comparison was made between the seroprevalence of ND antibodies in IV Department of Veterinary Pathology, Uni- versity of Ibadan, Nigeria household and live bird market as well as between sexes. Significantly higher prevalence rate (p<0.05) was observed with chickens sampled from households compared to those from the live bird market. Higher ranges of titers were also observed in chickens from households than those from live bird markets. The presence of ND antibodies in these unvaccinated local chickens and guinea fowls indicated the presence of the virus amongst the rural poultry population, hence aneed for improvement in vaccine campaignand delivery against ND for rural Mail Address poultry especially with the use of thermostable and improved oral or feed-based vaccine delivery systems. Corresponding author e-mail address Benjamin Emikpe Department of Pathobiology, School of Veterinary Medicine, Kwame Nkrumah INTRODUCTION University of Science and Technology, Kumasi, Ashanti, Ghana – 0233 Poultry production is the fastest growing component of global Phone: (+233) 549410841 meat production, with developing and transitional countries assuming Email: [email protected] a leading role (Assa, 2012). As indicated by Rojendran & Mohanty, (2003), poultry farming has become foremost among the subsidiary occupations of farmers to supplement their incomes because it assures Keywords quick returns, requires minimum space and investment, and can be Haemagglutination Inhibition test, Newcastle carried out by ordinary farmers. In Ghana, the poultry industry is an Disease, Vaccine, Kumasi, Ghana. important source of income and sustenance of livelihood for many individuals and families. Chicken eggs and meat are major sources of animal protein in the Ghanaian diet and their importance cannot be over-emphasized. This was reflected in the Veterinary Services Directorate (VSD) report for the year 2009, which showed that the poultry population of Ghana was slightly over 36.2 million, with local chicken forming 13.92% and guinea fowls 7.10% of the national total poultry population (VSD Annual reports, 2010). There is an increasing demand for chicken and chicken products in the Ghanaian market, which, coupled with an inadequate domestic production, determined Submitted: July/2015 a large volume of imports from other parts of the world. In 2010, Approved: October/2015 while domestic production was estimated in 37,279 tones of chicken 87 Boakye OD, Emikpe BO, Folitse RD, Bonnah SG, Adusei K, Owusu M, Serological Detection of Newcastle Disease Virus Oyebanji VO Antibodies in Local Chickens and Guinea Fowls in the Area of Kumasi, Ghana meat, the official figure of imported chicken meat was MATERIALS AND METHODS recorded as 72,418tonnes (VSD, 2011). However, local poultry industry in the country has been hampered Study area by numerous conditions including diseases, high cost The study was conducted in Kumasi Central Market of feed and medication, as well as competition with in the Kumasi Metropolis and Municipality, both cheaper imported frozen chicken products from the in the of Ghana. The Ashanti Region European markets (Aning, 2006) and elsewhere. includes one (1) Metropolitan, six (6) Municipal and Newcastle Disease has been identified as a major twenty (20) District Assemblies. The total land area is impediment to national poultry production particularly 24,389 square kilometers with the regions surrounding among the rural poultry population (Bensink & the Ashanti capital Kumasi having a population of Spradbow, 1999) in Ghana. Also, about80% of the 11 million Ashanti’s (Ajani & Ocobock 2012). The annual mortality of rural poultry has been attributed southern districts lies in thehumid semi-deciduous to outbreaks of Newcastle disease (VSD, 1998), which forest, whilst the Guinea savannah covers the northern are unpredictable and often discourage villagers from part. The administrative capital is Kumasi. The Kumasi paying proper attention to the husbandry and welfare Metropolitan area has an approximate area of 254 of their chickens (Alders & Spadbrow, 2000). In 2013, square kilometers and it is located between latitudes the annual report of the Veterinary Investigations 60035” and 6004”N and longitudes 10030” and 10035” Laboratory, Kumasi also recorded 97 different cases E. It shares boundaries with the Kwabre District to the of Newcastle disease,in which a total of 2,509 birds north, AtwimaKwanwoma and AtwimaNwabiagya diedand 334,006 of commercially-housed birds were District to the west, Ejisu- Municipal to the east exposed. However, there is dearth of information in and to the south (Ghana Statistical Ghana on the activity of the Newcastle disease virus Service, 2006). in rural poultry population and chickens from live The Ejisu-Juaben Municipality lies within Latitudes bird markets, which have been identified to be often 10 15’ N and 10 45’ N and Longitude 60 15’ W and involved in the spread of the disease (Spradbrow, 1988). 70 00’ W, occupying a land area of 582.5 km2. The The Government of Ghana recently launched poultry Municipality lies in central part of the Ashanti Region, projects aimed at boosting local poultry production sharing boundaries with six Districts in the Region. The and empowering livelihoods, especially in the rural district capital is sited at Ejisu (Ghana Statistical Service, communities where these projects are located, and 2014). therefore, the need to understand the ND virus activity Study design in rural poultry population in backyard and small-scale poultry farms is crucial. This investigation evaluates the Multistage sampling was carried out using activity of the Newcastle disease virus in local chicken stratified random sampling. The three regions and guinea fowl populations in live bird markets and where poultry populations are high in Ghana were households in Kumasi and its environs. listed. They are BrongAhafo, Ashanti, and Greater

Figure 1a: Map of Ghana Figure 1b: Map of Kumasi Figure 1c: Map of Ashanti Region

88 Boakye OD, Emikpe BO, Folitse RD, Bonnah SG, Adusei K, Owusu M, Serological Detection of Newcastle Disease Virus Oyebanji VO Antibodies in Local Chickens and Guinea Fowls in the Area of Kumasi, Ghana

Accra Region (Fig 1a,b,c). A random selection was RBCs were allowed to settle for about 30 minutes made andthe Ashanti Region was chosen. In the at room temperature. The agglutination is assessed Ashanti region, districts which also have some of by tilting the plates with those wells in which the the highest poultry populations were identified. RBCs streaming was observed at the same rate as They are Atwima-Nwabiagya, Kumasi, and Ejisu- the control wells (positive serum, virus/antigen and Juaben districts. Two districts were then randomly PBS controls) considered to show inhibition. selected, and the Kumasi and Ejisu-Juaben districts were chosen. In Kumasi, areas in which local fowls Data Analysis and guinea fowls are likely to be concentrated were Microsoft Excel 2010 spreadsheet was used to identified. The Kumasi Central Market was chosen. categorize the data and calculate the geometric mean The Kumasi Central Market is main destination for titers. The HI data obtained were submitted toanalysis commercial transactions of local chickens brought of variance using the SPSS package version 16. in from Kumasi city and other areas. In selecting the Categorical variables (sex, location, bird type, place of birds, a systematic random sampling was made, and purchase) were evaluated using the Chi-square test at th each bird on the 5 count was selected. 95% confidence interval to measure the strength of At Ejisu-Juaben district, the towns in which the association among variables. Values of p< 0.05 were local chickens were concentrated were identified considered significant. to be Besease, Juaben, Adumasa and Pokuase. A list of these townships was made and Besease Results and Adumasa townships were randomly selected. The interviews conducted revealed that the birds In the towns, a house was selected on each third were usually not vaccinated, as the last vaccination count and 3-5 birds were randomly selected by the reported by the farmers was in 2011. The interview householders themselves. A verbal interview was conducted. The selected birds were then sampled also showed that women and children were those taking 3-5 mL of blood from the wing vein using most involved in rural poultry production. It also a sterile 5 mL syringe, which was emptied into a revealed that farmers recognized the clinical features clean test tube and keptat room temperature for 5-6 (e.g. torticollis, high mortality, etc.) associated with ND hours. The tubes were then labeled appropriately, with periodic outbreaks often experienced during the indicating fowl type and sex. The sera were dry season. collected and stored under -20°Cin Eppendorf Table 1 shows birds from households presented tubes. Haemagglutination Inhibition test was then higher prevalence of ND than those collected in the performed using the OIE standard protocol. Briefly, market. In Ejisu- Besease, 83.3% females from tested 0.025 mL of PBS was dispensed into each well of positive while all (100%) males werepositive. In Ejisu- a plastic V-bottomed microliter plate then 0.025 Adumasa, the prevalence in females was higher mL of serum was placed into the first well of the (98.4%) than that recorded at Ejisu-Besease, while the plate. Two-fold dilutions of 0.025 mL serum were same situation (100%) was observed in males from the then made across the plate. ND viral antigen (0.025 two areas. Female guinea fowls presented a prevalence mL)was added to each well and the plate was left of 25.7%, while males had a prevalence of 22.9%. for 30 minutes at room temperature. Chicken Red However, the mean titers of the positive males were Blood Cells (RBCs) (0.025 mL of 1% (v/v))was then lower compared with those positive females, although added to each well and, after gentle mixing, the not statistically significant.

Table 1 – Prevalence of Newcastle disease antibodies in local chickens and guineafowls from various locations in Kumasi and its environs LOCATION Numbers of chickens Total Females(f) Males(m) F(+) Titer M(+) Titer Kumasi Central Market 57 24(42.1%) 33(57.9%) 12(50%) 5.2 6(18.2%) 5.6 Ejisu- BeseaseHousehold 85 72(84.7%) 13(15.3%) 60(83.3%) 6.3 13(100%) 7.7 Ejisu-AdumasaHousehold 150 126(84.0%) 24(16.0%) 124(98.4%) 9.8 24(100%) 11 Total 292 222(76.0%) 70 (24.0%) 196(88.3%) 8.2 43(61.4%) 9.0 Numbers of guineafowls Kumasi Central Market 153 70(45.8%) 83(54.2%) 18(25.7%) 6.9 19(22.9%) 5.4

89 Boakye OD, Emikpe BO, Folitse RD, Bonnah SG, Adusei K, Owusu M, Serological Detection of Newcastle Disease Virus Oyebanji VO Antibodies in Local Chickens and Guinea Fowls in the Area of Kumasi, Ghana

Table 2 – The ND titer ranges of local chicken and guinea fowls obtained from household and live bird markets in Kumasi, Ghana.

BIRD TYPE AREA OF ACQUISITION NEWCASTLE TITERRANGE (Log2) 0 1-5 6-10 ABOVE 10 Total Guinea fowl Market 116 12 21 4 153 75.8% 7.8% 13.7% 2.6% 100.0% Local chicken Market 40 6a 8c 3e 57 70.2% 10.5% 14.0% 5.3% 100.0% Local chicken Household 14 19b 74d 128f 235 6.0% 8.1% 31.5% 54.5% 100.0% p Value = 0.05, *values with different superscript indicates statistical difference (p<0.05)

Table 2 shows that the titer ranges of birds from In the latter,localchickens from households had higher the householdsaresignificantly different from those titers than those obtained inthe market. In the market, of from those obtained in themarket. It also showed 48.6% of guinea fowls were tested positive for ND, various titer ranges of both local chicken and guinea being 25.7% females and 22.9% males. This is in sharp fowls, where the birds that tested negative or had no contrastwith the existing reports of the live market asa detectable ND antibodies were designated 0, and those hub of ND viral activity (Jibril et al., 2014). The lower that tested positive were grouped 1-5, 6-10 and above ND activity in the live market in Kumasi could be due to

10 (Log2). Most of the local chickens from households lower ND viral activity in the birds sampled. presented titers higher than 6log2, whereas around The range of titers observed in birds from the live 70% of those in the market were negative. The guinea bird markets were similar, as most guinea fowls and fowls obtained in the market presented antibody titer local chickens presented titers between GMT 5-10, distribution similar to that of the local market chickens. while the mean titers of most of the local chickens sampled in households were higher 10. This suggests DISCUSSION that chickens from households possibly experience episodes of ND viral exposure and that the duration of This serological study revealed the presence of exposure is possiblylonger than those of the live bird circulating antibodies of Newcastle disease (ND) market. among local chickens and guinea fowls in Kumasi and its environs, with 81.8% of local chicken and 24.2% Conclusion of guinea fowls tested positive. This shows that ND is still an endemic viral disease of rural poultry in Ghana, This study shows that there is subclinical Newcastle as earlier observed by Awuni, (2002). ND prevalence Disease activity in rural poultry in Kumasi and its was significantly higher in females (hens) than males environs, which could easily escalate to disease state (cocks). This observation differs from a similar study in the event of stress. This is especially true with birds carried out in Nigeria by Jibril et al. (2014), who found from households, which possibly have repeated ND higher overall prevalence of ND in male chickens than virus infections, accounting the higher ND titer in these in females. This could possibly be due to the fact that birds compared with those sampled in the live-bird more hens were sampled in the present study. Males, market. Emphasis should therefore place on policies however, presented higher antibodies titers (GMT-9.0) to improve the vaccination campaign against ND in than females (GMT-8.2) possibly because the sampled rural poultry, particularly those in different households, males were adults, and this allows for multiple with the use of thermostable and better oral or in-feed exposures to ND virus in the environment. vaccine delivery systems. The ND prevalence inguinea fowls was different from that observed in the local chicken. The overall References prevalence was 24.2%, with females having a Ajani C, Ocobock P. Location: the ashanti lay roots in the Gold Coast. 2012. prevalence of 25.7% and males of22.9%. Although Available from: https://nd.digication.com/ajani_crosley/Ashanti. this indicates that there is some activity of the ND virus in guinea fowls, showing that, although significant, it Alders RG, Spradbrow PB. Newcastle disease in village chickens: a field manual. Proceedings of the SADC Workshop Newcastle Disease in is not as high as in the local chickens, as detected in Village Chickens; 2000 March 6-9; Maputo (Mozambique): ACIAR/Aus the present study and that of Oluwayelu et al. (2005). said; 2000.

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